Specification
Sheet
Please note
Some information
is Lot dependent,
Monoclonal Mouse
Anti-Human p21
WAF1/Cip1
Clone SX118
Code No,M 7202
Lot 088,Edition 09.11.00
Introduction p21 is a protein encoded by the WAF1/Cip1 gene on chromosome 6p,p21
WAF1/Cip1
has
been shown to inhibit the activity of several cyclin/cyclin-dependent kinase complexes and
to block cell cycle progression (1),It was identified as a gene whose product was
transcriptionally activated by wild-type but not mutant p53 protein,serving as a mediator of
the cell cycle arrest function of p53 (2),In tumour cells that have lost the p53 protein or
contain an altered form of p53,p21 levels are dramatically reduced or totally absent (2),
This could lead to the passing of the G1 checkpoint,permitting the progression of the cell
cycle in the presence of DNA alterations and in this way may result in genomic instability
and oncogenesis,Because p21 appears to mediate several of the growth-regulatory
functions of p53,its expression would be predicted to reflect the functional status of p53
more precisely than p53 accumulation,The p21
WAF1/Cip1
protein can also be induced in a
p53-independent manner and has been shown to play a role in differentiation (3),
Furthermore,p21
WAF1/Cip1
can inhibit DNA replication by blocking the action of proliferating
cell nuclear antigen (PCNA) (4),The expression of p21
WAF1/Cip1
has been studied by
immunohistochemistry in a wide range of human tumours including melanomas (5),
pancreatic carcinomas (6),cervical carcinomas (7),thymomas (8),thyroid carcinomas (9),
breast carcinomas (10),head-and-neck carcinomas (11),colon carcinomas (12) and
Hodgkin’s disease (13),
Presentation Monoclonal mouse antibody supplied in liquid form as tissue culture supernatant (RPMI
1640 medium containing fetal calf serum) dialysed against 0.05 mol/L Tris/HCl,pH 7.2
containing 15 mmol/L NaN
3
,
Mouse Ig concentration,390 mg/L,
Isotype,IgG1,kappa,
Total protein concentration,11.1 g/L,
Storage 2 - 8 °C,
Clone SX118,(14),
Immunogen Purified glutathione S-transferase (GST)-p21
WAF1/Cip1
fusion protein,
Specificity/reactivity The antibody reacts with a 21 kDa protein in immunoblotting of MCF-7 cells and
immunoprecipitates a 21 kDa band from lysates from SAOS-2 cells transfected with the
full length p21
WAF1/Cip1
protein (14),The antibody also reacts with p21 from mouse,
Application The antibody can be used in immunohistochemistry,Furthermore,the antibody produced
by the clone SX118 has been shown to be applicable for immunoblotting and
immunoprecipitation (14),
Staining procedures Formalin-fixed and paraffin-embedded sections
Can be used on formalin-fixed,paraffin-embedded tissue sections,Antigen retrieval,such as
by heating in 10 mmol/L citrate buffer,pH 6.0,or in DAKO Target Retrieval Solution,code
No,S 1700,is mandatory,The slides should not be allowed to dry out during this treatment
or during the following immunohistochemical staining procedure,
(2)
- 2 -
For tissue sections sensitive staining techniques are recommended,such as the LSAB?+ or
the EnVision?+ systems,
The antibody may be used at a dilution of 1:25 - 1:50 with the LSAB?+ system when tested
on formalin-fixed,paraffin-embedded sections of human tonsil,
Frozen sections and cell smears
Can be used for labelling acetone-fixed cryostat sections or for fixed cell smears,For
staining cell smears,the APAAP method is recommended,
The antibody may be used at a dilution at 1:25 - 1:50 in the APAAP technique,when tested
on acetone-fixed cryostat sections of human tonsil,
These are guidelines only; optimal dilutions should be determined by the individual
laboratory,
Automation,The antibody can be used on automated immunostaining systems,
References (1) Xiong Y,Hannon GJ,Zhang H,Casso D,Kobayashi R,Beach D,p21 is a universal inhibitor of cyclin
kinases,Nature 1993;366:701-4,
(2) El-Deiry WS,Tokino T,Velculescu VE,Levy DB,Parsons R,Trent JM,et al,WAF1,a potential mediator
of p53 tumor suppression,Cell 1993;75:817-25,
(3) Steinman RA,Hoffman B,Iro A,Guillouf C,Liebermann DA,El-Houseini ME,Induction of p21 (WAF-
1/CIP1) during differentiation,Oncogene 1994;9:3389-96,
(4) Li R,Waga S,Hannon GJ,Beach D,Stillman B,Differential effects by the p21 CDK inhibitor on PCNA-
dependent DNA replication and repair,Nature 1994;371:534-7,
(5) Gorospe M,Cirielli C,Wang X,Seth P,Capogrossi MC,Holbrook NJ,p21
Waf1/Cip1
protects against p53-
mediated apoptosis of human melanoma cells,Oncogene 1997;14:929-35,
(6) Dergham ST,Dugan MC,Joshi US,Chen YC,Du W,Smith DW,et al,The clinical significance of
p21
WAF1/CIP1
and p53 expression in pancreatic adenocarcinoma,Cancer 1997;80:372-81,
(7) Palazzo JP,Mercer WE,Kovatich AJ,McHugh M,Immunohistochemical localization of p21
WAF1/CIP1
in
normal,hyperplastic,and neoplastic uterine tissues,Hum Pathol 1997;27:60-6,
(8) Stefanaki K,Rontogianni D,Kouvidou CH,Bolioti S,Delides G,Pantelidaki A,et al,Expression of p53,
mdm2,p21/waf1 and bcl-2 proteins in thymomas,Histopathol 1997;30:549-55,
(9) Ito Y,Kobayashi T,Takeda T,Komoike Y,Wakasugi E,Tamaki Y,et al,Expression of p21 (WAF1/CIP1)
protein in clinical thyroid tissues,Br J Cancer 1996;74:1269-74,
(10) Guillot C,Falette N,Paperin M-P,Courtois S,Gentil-Perret A,Treilleux I,et al,p21
WAF1/CIP1
response to
genotoxic agents in wild-type TP53 expressing breast primary tumours,Oncogene 1997;14:45-52,
(11) Erber R,Klein W,Andl T,Enders C,Born AI,Conradt C,et al,Aberrant p21
CIP1/WAF1
protein accumulation
in head-and-neck cancer,Int J Cancer 1997;74:383-9,
(12) Valassiadou KE,Stefanaki K,Tzardi M,Datseris G,Georgoulias V,Melissas J,et al,
Immunohistochemical expression of p53,bcl-2,mdm2 and waf1/p21 proteins in colorectal
adenocarcinomas,Anticancer Res 1997;17:2571-6,
(13) Naresh KN,O’Conor GT,Soman CS,Johnson J,Advani SH,Magrath IT,et al,A study of p53 protein,
proliferating cell nuclear antigen,and p21 in Hodgkin’s disease at presentation and relapse,Hum Pathol
1997;28:549-55,
(14) Fredersdorf S,Milne AW,Hall PA,Lu X,Characterization of a panel of novel anti-p21
Waf1/Cip1
monoclonal
antibodies and immunochemical analysis of p21
Waf1/Cip1
expression in normal human tissues,Am J
Pathol 1996;148:825-35,
M 7202/JA/09.11.00